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Main Title: Quantification of Major Bacteria and Yeast Species in Kefir Consortia by Multiplex TaqMan qPCR
Author(s): Nejati, Fatemeh
Junne, Stefan
Kurreck, Jens
Neubauer, Peter
Type: Article
Abstract: Kefir grains are complex microbial systems of several groups of microorganisms. The identification and quantification of the microbial composition of milk kefirs was described in several studies, which provided an insight into the microbial consortia in this complex ecosystem. Nevertheless, the current methods for identification and quantification are not appropriate for deeper studies on kefir consortia, e.g., population dynamics and microbial interactions in kefir grains. This requires another sensitive and reliable quantitative method. Therefore, this study aims to develop multiplexed qPCR assays to specifically detect and quantify, as an example, several microorganisms of the milk kefir microbial community. Primer-probe sets, which target species-specific genes in six bacteria and five yeasts, were designed, and their sensitivity and specificity to the target species was analyzed in simplex as well as four multiplex qPCR assays. The self-designed multiplex assays were applied for the detection of target bacteria and yeast species in milk kefirs, in both, grain and beverage fractions. Detection of all target microorganisms in simplex and multiplex qPCR was achieved by good linearity, efficiency, repeatability and reproducibility in all assays. When the designed assays were applied on six kefirs, all target microorganisms were detected in different samples, but not all in one kefir sample. The two ubiquitous lactobacilli Lactobacillus kefiranofaciens and Lb. kefiri were present in all six kefirs studied, but were associated with different other yeasts and bacteria. Especially on the yeast community a significant diversity was observed. In general, multiplex TaqMan qPCR as developed here was proven to have high potential for specific identification of target microorganisms in kefir samples and for the first time, eleven target bacteria and yeasts of kefir microbiota were rapidly detected and quantified. This study, thus, provides a fast and reliable protocol for future studies on kefir and other similar microbial ecosystems.
Subject(s): TaqMan qPCR
milk kefir
microbial community
lactic acid bacteria
Issue Date: 16-Jun-2020
Date Available: 28-Aug-2020
Language Code: en
DDC Class: 570 Biowissenschaften; Biologie
Sponsor/Funder: DFG, 414044773, Open Access Publizieren 2019 - 2020 / Technische Universität Berlin
Journal Title: Frontiers in Microbiology
Publisher: Frontiers Media S.A.
Volume: 11
Article Number: 1291
Publisher DOI: 10.3389/fmicb.2020.01291
EISSN: 1664-302X
TU Affiliation(s): Fak. 3 Prozesswissenschaften » Inst. Biotechnologie » FG Bioverfahrenstechnik
Appears in Collections:Technische Universität Berlin » Publications

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